College of Wildlife and Protected Area, Northeast Forestry University, Harbin, China.
Department of Forest Protection, College of Forestry, Northeast Forestry University, Harbin, China; State Key Laboratory of Rice Biology & Ministry of Agriculture and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Insect Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.
Pestic Biochem Physiol. 2024 Feb;199:105765. doi: 10.1016/j.pestbp.2023.105765. Epub 2024 Jan 2.
The detoxification of insecticides in insects is dependent on the expression and activity of multiple detoxification enzymes. As an important modulator of detoxification enzymes, the CncC-Keap1 pathway was involved in the detoxification of various pesticides. However, whether the CncC-Keap1 pathway is involved in the detoxification of emamectin benzoate (EMB) is unclear. In this study, we cloned the LdCncC and LdKeap1 from spongy moths (Lymantria dispar). Our results showed that EMB exposure induced oxidative stress, and activated the CncC-Keap1 pathway at mRNA and protein levels. Removing ROS by N-acetylcysteine remarkably decreased HO levels and restored the expression of LdCncC and LdKeap1. The silencing LdCncC, not LdKeap1, by dsRNA significantly decreased the cytochrome P450 activities, and increased the sensitivity of larvae to EMB. Besides, the expression of CYP6B7v1, CYP321A7 and CYP4S4v1 were significantly decreased after silencing LdCncC. Notably, the knockdown of CYP6B7v1, CYP321A7 or CYP4S4v1 significantly increased the mortality induced by EMB exposure. Therefore, we proposed that activation of CncC-Keap1 pathway induced by ROS increased the detoxification of EMB in spongy moths by regulating the expression of CYP6B7v1, CYP321A7 and CYP4S4v1. Our study strengthened the understanding of the detoxification of EMB from the perspective of CncC-Keap1-P450s pathway.
昆虫中的杀虫剂解毒依赖于多种解毒酶的表达和活性。作为解毒酶的重要调节剂，CncC-Keap1 途径参与了各种杀虫剂的解毒。然而，CncC-Keap1 途径是否参与了甲氨基阿维菌素苯甲酸盐（EMB）的解毒尚不清楚。在本研究中，我们从透翅蛾（Lymantria dispar）中克隆了 LdCncC 和 LdKeap1。我们的结果表明，EMB 暴露诱导氧化应激，并在 mRNA 和蛋白质水平上激活 CncC-Keap1 途径。N-乙酰半胱氨酸去除 ROS 可显著降低 HO 水平并恢复 LdCncC 和 LdKeap1 的表达。dsRNA 沉默 LdCncC（而非 LdKeap1）可显著降低细胞色素 P450 活性，并增加幼虫对 EMB 的敏感性。此外，沉默 LdCncC 后 CYP6B7v1、CYP321A7 和 CYP4S4v1 的表达显著降低。值得注意的是，沉默 CYP6B7v1、CYP321A7 或 CYP4S4v1 后，EMB 暴露引起的死亡率显著增加。因此，我们提出 ROS 诱导的 CncC-Keap1 途径的激活通过调节 CYP6B7v1、CYP321A7 和 CYP4S4v1 的表达增加了透翅蛾对 EMB 的解毒。我们的研究从 CncC-Keap1-P450s 途径的角度加强了对 EMB 解毒的理解。
